In vivo imaging of neural reactive plasticity after laser axotomy in cerebellar cortex

A. L. Allegra Mascaro; L. Sacconi; B. Maco; G. W. Knott; F. S. Pavone

doi:10.1117/12.2037276

24 March 2014 In vivo imaging of neural reactive plasticity after laser axotomy in cerebellar cortex

A. L. Allegra Mascaro, L. Sacconi, B. Maco, G. W. Knott, F. S. Pavone

Proceedings Volume 8928, Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics; 89281N (2014) https://doi.org/10.1117/12.2037276
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

Multi-photon imaging provides valuable insights into the continuous reshaping of neuronal connectivity in live brain. We previously showed that single neuron or even single spine ablation can be achieved by laser-mediated dissection. Furthermore, single axonal branches can be dissected avoiding collateral damage to the adjacent dendrite and the formation of a persistent glial scar. Here, we describe the procedure to address the structural plasticity of cerebellar climbing fibers by combining two-photon in vivo imaging with laser axotomy in a mouse model. This method is a powerful tool to study the basic mechanisms of axonal rewiring after single branch axotomy in vivo. In fact, despite the denervated area being very small, the injured axons consistently reshape the connectivity with surrounding neurons, as indicated by the increase in the turnover of synaptic boutons. In addition, time-lapse imaging reveals the sprouting of new branches from the injured axon. Newly formed branches with varicosities suggest the possible formation of synaptic contacts. Correlative light and electron microscopy revealed that the sprouted branch contains large numbers of vesicles, with varicosities in the close vicinity of Purkinje dendrites.

Citation Download Citation

A. L. Allegra Mascaro, L. Sacconi, B. Maco, G. W. Knott, and F. S. Pavone "In vivo imaging of neural reactive plasticity after laser axotomy in cerebellar cortex", Proc. SPIE 8928, Optical Techniques in Neurosurgery, Neurophotonics, and Optogenetics, 89281N (24 March 2014); https://doi.org/10.1117/12.2037276

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KEYWORDS

In vivo imaging

Axons

Neurons

Californium

Electron microscopy

Dendrites

Optical fibers

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